Highly Selective Separation Of Gastrodin By SiO₂/Rosin-based Core-shell Liquid Chromatography Stationary Phase

Gastrodin is one of the effective active ingredients of traditional Chinese medicine Tianma.It is widely used in the field of biomedicine.At present,gastrodin is separated and purified by the petroleum-based macroporous resin stationary phase,which may leave toxic benzene-based substances and threate to human health in the drug during separation and purification.Rosin is a rich renewable biomass resource,which is favored by more and more people with its green environmental protection,unique ternary pheno-ring structure and good mechanical rigidity.In this thesis,SiO₂/rosin-based core-shell stationary phases were prepared by using rosin modified cross-linking agent as raw material to separate gastrodin and its derivatives.The main contents are as follows:1.Four kinds of SiO₂/rosin-based polymer microspheres were prepared by silica gelcoatingandusedasstationaryphaseinhigh-performanceliquid chromatography?HPLC?,named SP₁,SP₂,SP₃ and SP_4.In the preparation process,Acrylpimaric Acid Ethylene Glyyol Acrylate?AAEGA?and methyl methacrylate?MMA?were used as the cross-linker and monomer,respectively.SiO₂ microspheres were named SP₀.SiO₂ and SiO₂/rosin-based core-shell stationary phases were characterized by scanning electron microscope?SEM?,transmission electron microscope?TEM?,laser granulometry,specific surface area and pore structure analysis,thermogravimetric analysis.The results showed that the SiO₂/rosin-based core-shell stationary phases had good sphericity,the particle size distribution was in the range of 2-10?m,the specific surface area was between 275-305 m²/g,and the average pore diameter was between 6.31-6.36 nm.Decomposition started at 250°C,indicating it had good thermal stability.2.The SiO₂/rosin-based polymer microspheres and were filled into a stainl ess steel column by wet packing to obtain SP₁,SP₂,SP₃ and SP₄ column to s eparate the gastrodin and its derivatives?Phenyl-?-D-glucopyranoside and 4-meth oxyphenyl-?-D-glucopyranoside?.The influencing facts including concentration o f acetonitrile and phosphoric acid,flow rate and column temperature were expl ored.The results indicated that the SP₃ column had the best separation effect on gastrodin and its derivatives.The resolution of gastrodin and phenyl-?-D-glu copyranoside,gastrodin and 4-methoxyphenyl-?-D-glucopyranoside was 8.52 and14.7 when the mobile phase was acetonitrile-0.05%phosphoric acid?3:97 v/v?,t he temperature was 25°C,and the flow rate was 0.3 mL/min.3.The thermodynamics of the separation of gastrodin and its derivatives was explored by SP₃ column.The?H,?S,?G were calculated by the Van't Hoff equation.The results showed that separation of gastrodin and phenyl-?-D-glucopyranoside,gastrodin and 4-methoxyphenyl-?-D-glucopyranoside by the SP₃ column was both enthaply and entropy driven.4.Gastrodin was separated and purified by the SP₃ column form the aqueous extract of gastrodin elata.The results showed that the purity of gastrodin increased from 47.7%to 86.4%after separation and purification.